Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs
Human embryonic stem (hES) cells are an attractive source of cellular material for scientific, diagnostic, and potential therapeutic applications. Protocols are now available to direct hES cell differentiation to specific lineages at high purity under relatively defined conditions; however, resear...
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
Mary Ann Liebert
2011
|
| Online Access: | http://hdl.handle.net/1721.1/61718 |
| _version_ | 1826197998639513600 |
|---|---|
| author | Metallo, Christian M. Azarin, Samira M. Moses, Laurel E. Ji, Lin Pablo, Juan J. de Palecek, Sean P. |
| author2 | Massachusetts Institute of Technology. Department of Chemical Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Chemical Engineering Metallo, Christian M. Azarin, Samira M. Moses, Laurel E. Ji, Lin Pablo, Juan J. de Palecek, Sean P. |
| author_sort | Metallo, Christian M. |
| collection | MIT |
| description | Human embryonic stem (hES) cells are an attractive source of cellular material for scientific, diagnostic, and
potential therapeutic applications. Protocols are now available to direct hES cell differentiation to specific lineages
at high purity under relatively defined conditions; however, researchers must establish the functional
similarity of hES cell derivatives and associated primary cell types to validate their utility. Using retinoic acid to
initiate differentiation, we generated high-purity populations of keratin 14þ (K14) hES cell-derived keratinocyte
(hEK) progenitors and performed microarray analysis to compare the global transcriptional program of hEKs
and primary foreskin keratinocytes. Transcriptional patterns were largely similar, though gene ontology analysis
identified that genes associated with signal transduction and extracellular matrix were upregulated in hEKs. In
addition, we evaluated the ability of hEKs to detect and respond to environmental stimuli such as Ca2þ, serum,
and culture at the air–liquid interface. When cultivated on dermal constructs formed with collagen gels and
human dermal fibroblasts, hEKs survived and proliferated for 3 weeks in engineered tissue constructs. Maintenance
at the air–liquid interface induced stratification of surface epithelium, and immunohistochemistry results
indicated that markers of differentiation (e.g., keratin 10, involucrin, and filaggrin) were localized to
suprabasal layers. Although the overall tissue morphology was significantly different compared with human
skin samples, organotypic cultures generated with hEKs and primary foreskin keratinocytes were quite similar,
suggesting these cell types respond to this microenvironment in a similar manner. These results represent an
important step in characterizing the functional similarity of hEKs to primary epithelia. |
| first_indexed | 2024-09-23T10:57:17Z |
| format | Article |
| id | mit-1721.1/61718 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:57:17Z |
| publishDate | 2011 |
| publisher | Mary Ann Liebert |
| record_format | dspace |
| spelling | mit-1721.1/617182022-09-27T16:09:05Z Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs Metallo, Christian M. Azarin, Samira M. Moses, Laurel E. Ji, Lin Pablo, Juan J. de Palecek, Sean P. Massachusetts Institute of Technology. Department of Chemical Engineering Metallo, Christian M. Metallo, Christian M. Human embryonic stem (hES) cells are an attractive source of cellular material for scientific, diagnostic, and potential therapeutic applications. Protocols are now available to direct hES cell differentiation to specific lineages at high purity under relatively defined conditions; however, researchers must establish the functional similarity of hES cell derivatives and associated primary cell types to validate their utility. Using retinoic acid to initiate differentiation, we generated high-purity populations of keratin 14þ (K14) hES cell-derived keratinocyte (hEK) progenitors and performed microarray analysis to compare the global transcriptional program of hEKs and primary foreskin keratinocytes. Transcriptional patterns were largely similar, though gene ontology analysis identified that genes associated with signal transduction and extracellular matrix were upregulated in hEKs. In addition, we evaluated the ability of hEKs to detect and respond to environmental stimuli such as Ca2þ, serum, and culture at the air–liquid interface. When cultivated on dermal constructs formed with collagen gels and human dermal fibroblasts, hEKs survived and proliferated for 3 weeks in engineered tissue constructs. Maintenance at the air–liquid interface induced stratification of surface epithelium, and immunohistochemistry results indicated that markers of differentiation (e.g., keratin 10, involucrin, and filaggrin) were localized to suprabasal layers. Although the overall tissue morphology was significantly different compared with human skin samples, organotypic cultures generated with hEKs and primary foreskin keratinocytes were quite similar, suggesting these cell types respond to this microenvironment in a similar manner. These results represent an important step in characterizing the functional similarity of hEKs to primary epithelia. National Institute of Biomedical Imaging and Bioengineering (U.S.) (Grant 1R01EB007534) National Science Foundation (U.S.) (Grant EFRI-0735903) National Institutes of Health (U.S.). Biotechnology Training Fellowship 2011-03-17T22:08:08Z 2011-03-17T22:08:08Z 2010-01 Article http://purl.org/eprint/type/JournalArticle 1937-3341 1937-335X http://hdl.handle.net/1721.1/61718 Metallo, Christian M. et al. “Human Embryonic Stem Cell-Derived Keratinocytes Exhibit an Epidermal Transcription Program and Undergo Epithelial Morphogenesis in Engineered Tissue Constructs.” Tissue Engineering Part A 16.1 (2010): 213-223. c2010 Mary Ann Liebert en_US http://dx.doi.org/10.1089/ten.TEA.2009.0325 Tissue engineering. Part A Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Mary Ann Liebert Mary Ann Liebert |
| spellingShingle | Metallo, Christian M. Azarin, Samira M. Moses, Laurel E. Ji, Lin Pablo, Juan J. de Palecek, Sean P. Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title | Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title_full | Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title_fullStr | Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title_full_unstemmed | Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title_short | Human embryonic stem cell-derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| title_sort | human embryonic stem cell derived keratinocytes exhibit an epidermal transcription program and undergo epithelial morphogenesis in engineered tissue constructs |
| url | http://hdl.handle.net/1721.1/61718 |
| work_keys_str_mv | AT metallochristianm humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs AT azarinsamiram humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs AT moseslaurele humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs AT jilin humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs AT pablojuanjde humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs AT palecekseanp humanembryonicstemcellderivedkeratinocytesexhibitanepidermaltranscriptionprogramandundergoepithelialmorphogenesisinengineeredtissueconstructs |